Investigation of conserved Flagellum proteins in Trypanosoma brucei

The single celled protozoan parasite Trypanosoma brucei is an excellent model organism to study eukaryotic cilia and flagella as it has a single flagellum that remains assembled throughout the cell cycle. The new flagellum assembles in a known position relative to the old flagellum, therefore creating a model system of identifiable organelle generations. In additional to a sequenced genome, there are many reverse genetics tools developed for T. brucei which makes the functional analysis of proteins possible. More than 300 proteins have been identified as components of the T. brucei flagellum but functional analysis of the majority of these proteins has not been carried out to date. This project used a bioinformatics approach to identify potential flagellum proteins in T. brucei that were also conserved in Homo sapiens, thereby identifying potential ciliopathy candidates. Candidate proteins were confirmed as flagellum components through endogenous localisation techniques and co-localisation studies. Functional analysis was performed using inducible RNAi cell lines. Light and electron microscopy techniques were used for phenotypic analysis. Through bioinformatics analysis a novel family of coiled-coil TPH domain-containing proteins were identified that are highly conserved in flagellated eukaryotes. There are three TPH domain-containing proteins conserved in T. brucei that all have a role in flagellum length control and cell morphogenesis. In all three cases protein ablation has a detrimental effect on cellular motility. This work provides further understanding into the complexities of flagellum biogenesis in T. brucei and the downstream effects on cell motility and morphogenesis.

Grantor dc:publisher

Oxford Brookes University

Year dc:date

2015

Author dc:creator

• Borrett, Samantha J.

Contributors dc:contributor

• Vaughan, Sue

Statement dc:rights

• All rights reserved

Language dc:language

en

DOI dc:identifier

https://doi.org/10.24384/wgxt-ka37

OAI identifier oai:identifier

tle:a85a4fc5-4ecf-41ea-a385-51a546281800:d6bd9758-527a-46cd-bfe2-c433766e8fca:1